MID-TERM STORAGE AND GENETIC STABILITY OF STRAWBERRY TISSUE CULTURES
Abstract
The objectives of the present investigation were to establish an efficient medium-term in vitro preservation protocol for strawberry and to assess the genetic stability of the stored cultures. The medium term preservation study was initiated under 4°C and dark conditions using in vitro grown shootlet cultures. Shoot explants were subjected to different concentrations of mannitol and sorbitol (0.1, 0.2 and 0.4 M) as osmotic regulators supplemented to the culture medium which contained or was devoid of Indolbutyric acid (IBA). Results showed that the number of shoots increased when the culture medium was supplemented with 1 mg IBA/L and 0.1 M sorbitol and preserved for 15 months. However, the highest percentage of healthy shoots (76%) was observed with the medium containing 0.2 M sorbitol without IBA. Whereas, the highest number of roots per shoot occurred on the medium supplemented with 0.1 M sorbitol devoid of IBA. Also, sorbitol containing medium registered best results of shootlet recovery. Determination of stability was performed by assessment of randomly amplified polymorphic DNA (RAPD). The total number of amplified RAPD fragments varied depending on the sequence of the primers used. The generated RAPD profiles show no variation among the preserved and nonpreserved material with all primers tested.References
Bekheet, S. A. (2007). In vitro preservation of Globe artichoke germplasm. Plant Tissue Cult. & Biotech., 17: 1-9.
Brown, P. H., F. D. Lange, E. Kranz and H. Lorz (1993). Analysis of single protoplasts and regenerated plants by PCR and RAPD technology. Mol.Gen.Genet., 237: 311-317.
Cecchini, E., N. Natali, A. Cavallini and M. Durante (1992). DNA variation in regenerated plants of pea (Pisum sativum L.). Theor. Appl. Genet., 84: 874-879.
Dekkers, A. J., A. N. Rao and C. J. Goh (1991). In vitro storage of multiple shoot cultures of ginger at ambient temperature. Sci. Hortic., 47: 157- 167.
Dodds, J. H. and W. Roberts. (1985). Experiments In Plant Tissue Culture. 2nd Edition. Cambridge University Press, Cambridge, UK.
Engelmann, F. (1997). In vitro conservation methods. Pp. 119-162 in Biotechnology and Plant Genetic Resources: Conservation and Use. (J.A. Callow, B.V. Ford-Lloyd and H. J. Newbury (eds). CAB International, Wallingford, UK. Espinoza, N., R. Esterada, P. Tovar, J.
Bryan and J. H. Doddas (1984). Tissue culture micro propagation conservation and export of potato germplasm. Specialized Technology Document 1, CIP, Lima. Peuro
Espinoza, R A., H. L. Salas, P. O. Gonzalez and P. J. J Silva (2002). Use of abscisic acid, mannitol and salt concentration decrease in the culture medium in the in vitro conservation of Ipomoea batatas. Biotecnologyia- Vegetal., 2: 39-42.
Flecher, P. J. (1994). In vitro long-term storage of asparagus: New Zealand J. Crop Hort. Sci., 22: 351-359.
Graham, J., R. J. McNicol, K. Greig, and W. T. G. Van de Ven (1994). Identification of red raspberry cultivars and an assessment to their relatedness using fingerprintings produced by random primers. J. Hort. Sci., 69: 123-130.
Gupta, S. (2001). ABA-induced growth retardation of kiwifruit (Actinidia chinensis) in vitro. Indian J. Plant Genet. Resource, 14: 312-313.
Hae Boong J., H. SsangHee and K. K. Yoon (1996). In vitro Preservation method of culturing shoot tip at low temperature in strawberry. RDA Journal of Agriculture Science Biotechnology, 38: 284- 289.
Jarret, R. L. and N. Gawel (2004). Chemical and environmental growth regulation of sweetpotato (Ipomoea batatas L.) Lam. in vitro. Plant Cell, Tissue and Orange Culture. Pp. 153-159.
Lanham, P. G. and R. M. Brennan (1998.). Characterization of genetic resources of redcurrant (Ribes rubrum Ribesia) using anchored microsatellite markers. Theor. Appl. Genet., 96: 917-921. Lundergan, C. and J. Janick (1979). Low temperature storage of in vitro apple shoot. Hort. Science, 14: 514.
Mandal, B. B. (1995). Methods of in vitro conservation: principles, prospects and constraints. Pp. 83-87 in Plant Germplasm Conservation: Biotechnological Approaches. (R.S. Rana, K.P.S. Chandel, S. R. Bhat, B. B. Mandal, J. L. Karihaloo, K. V. Bhat and R. Pandey, eds.). NBPGR, New Delhi, India.
Mandal, B. B. (1999). Conservation biotechnology of endemic and other economically important plant species of India. Pp. 211-225 in Plant Conservation Biotechnology. (E.E. Benson, ed.). Taylor and Francis, London, UK.
Monette, P. L. (1986) .Cold storage of kiwi fruit shoot tips in vitro. Hort. Science, 11: 1203-1205.
Morel, G. (1975). Merstim culture techniques for the long-term storage of cultivated plants, P. 327-332. In: O. H. Frankel and J. G. Hawkes (eds.). Crop Genetic Resources for Today and Tomorrow. Cambridge Univ. Press, London
Moreno, S., Y. Gogorcena and J. M. Ortiz (1995). The use of RAPD markers for identification of cultivated grapevine (Vitis vinifera L.). Sci. Hort., 62: 237-243.
Mulis, K. B. and F. A. Faloona (1987). Specific synthesis of DNA in vitro via a polymerase-catalysed chain reaction. Methods Enzymol., 155: 335-350.
Mullen, R. H. and D. E. Schlegel (1976). Cold storage maintenance of strawberry meristems plantlets. Hort. Science, 11: 100-101.
Ng, SYC and S. K. Hahan (1985). Application of tissue culture to tuber crops at IITA. In: Proceeding of the Inter-Center Seminar on International Agricultural Research Centers and Biotechnology. IRRI, Philippines, pp27-40.
Reed, B. M. (1991). Application of gaspermeable bags for in vitro cold storage of strawberry germplasm. Plant Cell Rep., 10: 431-434.
Reed, B. M. (1992). Cold storage of strawberries in vitro: a comparison of three storage systems. Fruit Varieties Journal, 46: 98-102.
Reed, B. M. and K. E. Hummer (1995). Conservation of Germplasm of Strawberry (Fragaria Species) In: Y.P.S. Bajaj (Ed.) Biotechnology in Agriculture and Forestry, 32: 354-370.
Reed, B. M. (2002). Photoperiod improves long-term survival of in vitro-stored strawberry plantlets. Hort. Science, 37: 811-814.
Tompson, M. R., T. Douglas, J. Obatasamoto and T. A. Thorpe (1986). Mannitol metabolism in cultured plant cells. Physiol. Plant, 67: 365- 369.
Sabir, A., H. J. Newbury, G. Todd, J. Catty and B. V. Ford-Lloyd (1992). Determination of genetic stability using isozymes and RFLP in beet plants regenertated in vitro. Theor. Appl. Genet., 84: 113-117.
Saiki, R. K., S. Scharf, F. Faloona, K. B. Mullis, Horn, G. T. H. A. Erlich and N. A. Arnheim (1985). Enzymatic amplification of globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science, 230: 1350- 1354.
Waller, R. A. and D. B. Duncan (1969). A basic rule for thaw symmetric multiple comparison problems. Amer. States Assoc. J. Dec.: 1485- 1503.
Westcott R. J. (1981). Tissue culture storage of potato germplasm 2. Use of growth retardants. Potato Res. 24:343-352.
Williams, J. K. G., A. R. Kubelik, K. J. Livak, J. A. Rafalski and S.V. Tingey (1990). DNA polymorphism amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res., 18: 6531-6535.
Withers, L. A. (1984). Germplasm conservation in vitro: present state of research and its application. Pp. 138-157 in Crop Genetic Resources: Conservation and Evaluation. J. H. W. Holden and J. T. Williams (eds.). George Allen and Unwin, London, UK. Withers, L. A. (1987). Long-term preservation of plant cell, tissues and organs. Oxford Surv. Plant Mol. Cell. Biol., 4: 221-272.
Withers, L. A. (1991). Biotechnology and plant genetic resources conservation. In Paroda, R. S., Arora R. K. (ed.): Plant Genetic Resources, Conservation and Management Concepts and Approaches Pp. 273-297. International Board for Plant Genetic Resources, New Delhi.
Withers, L. A. and F. Engelmann (1998). In vitro conservation of plant genetic resources. Pp. 57-88 in Biotechnology in Agriculture. (A. Altman, ed.). Marcel Deckker Inc., New York, USA.
Yu Gui Hong, W., Z. WeiMin, L. Mizhen and Q. Y. XiaoGang (2003). Experiment of low temperature preservation of strawberry genetic resources. South China Fruits, 32: 54.
Zamora, A. B. and C. N. Paet (1999). In vitro genebank activities at the institute of plant breeding, College of Agriculture, University of the Philippines at Los Banos. Pp. 76- 83 in Management of field and in vitro germplasm collections. Proceedings of a Consultation Meeting, 15-20 January 1996, CIAT, Cali, Colombia. (F. Engelmann, ed.). IPGRI, Rome, Italy. Table
